水稻条纹叶枯病毒RNA干扰载体的构建及遗传转化  

Construction and Genetic Transformation of RNA Interference Vectors for Rice Stripe Virus

作　　者：李海青[1,2,3] 柳絮[2,3] 李臻[2,3] 姚方印[2,3] 刘炜[1,2,3] 

机构地区：[1]山东师范大学生命科学学院,山东济南250014 [2]山东省农业科学院高新技术研究中心/山东省作物遗传改良与生物技术重点实验室,山东济南250100 [3]农业部黄淮海作物遗传改良与生物技术重点开放实验室,山东济南250100

出　　处：《山东农业科学》2011年第2期1-6,共6页Shandong Agricultural Sciences

基　　金：国家转基因植物新品种培育重大专项(编号:2009ZX08001-010B)资助

摘　　要：利用已知的水稻条纹叶枯病毒的序列设计了12对引物,利用这些引物经RT-PCR获得了5条序列,经BLAST分析,它们均与水稻基因组内的序列完全不匹配,可以用来作为干扰序列。将这5个序列构建入由pUC19改造的一段内含子两侧各含有一对同尾酶的干扰载体中,然后再将整个RNA干扰片段构建入经改造的以玉米泛素Ubi为启动子、以生物安全性的bar基因为选择标记的植物双元表达载体pCAMBIA1300中,经农杆菌介导对黄淮稻区主栽水稻品种圣稻13进行遗传转化,为抗条纹叶枯病水稻新品种的培育提供了候选材料。Twelve pairs of primers were designed according to the known sequences of rice stripe virus,and 5 sequences were obtained by RT-PCR.By BLAST analysis,no similar sequences in rice genome were found matching these five sequences,so they could be used as the interference sequences.A modified pUC19 vector inserted a fragment as intron with a pair of isocaudarners on each side was used as the interference vector.After inserting the interference sequences into the interference vector separately,the complete interference fragments were integrated into the modified binary vector pCAMBIA1300 using ubiquitin as promoter and bar gene as selectable marker.The transgenic rice seedlings were obtained by Agrobacterium-mediated transformation to rice variety Shengdao 13.This provided candidate materials for breeding new rice varieties resistant to rice stripe virus.

关 键 词：水稻条纹叶枯病毒 RNA干扰载体 Ubi启动子 BAR基因 抗病水稻品种 

分 类 号：Q782[生物学—分子生物学]